Detergent Removal from Peptides | Thermo Fisher Scientific

The Thermo Scientific Pierce Detergent Removal Resins are provided in convenient spin-column or plate formats that quickly and efficiently remove ionic, nonionic, and/or zwitterionic detergents from protein or peptide samples to improve compatibility with downstream applications. Two formulations are available that are optimized to remove detergents from peptide samples with different concentration ranges. The Thermo Scientific HiPPR (high protein and peptide recovery) products are recommended for peptide samples ≤100 μg/mL. The standard Pierce Detergent Removal Resin products are ideal for peptide samples >100 μg/mL.

High performance—removes detergent with >90% recovery and no sample dilution
Versatile—effectively removes a wide variety of detergents from peptide or protein samples
Optimized—separate formulations for samples with peptide concentrations ≤ or >100 μg/mL
Flexible—available in various formats, including spin columns, 96-well spin plates, and loose resin
Convenient—simple method that helps to improve MS peptide coverage

Choose the right peptide detergent removal product for your application

	HiPPR Detergent Removal Spin Columns	HiPPR Detergent Removal Spin Kit	HiPPR Detergent Removal Spin Plates	Detergent Removal Spin Columns (4 sizes)	Detergent Removal Resin	Detergent Removal Spin Plates

Sample volume range(s)	100 µL	25– 200 µL	100 µL	10– 25 µL 25– 100 µL 150– 500 µL 500– 1,000 µL	10– 2,500 µL	20– 100 μL
Recommended peptide sample concentration range	1– 100 µg/mL	1–100 µg/mL	1–100 µg/mL	>100 µg	>100 µg	>100 µg
Format	Pre-filled spin columns	5 mL resin + 0.8 mL empty spin columns	96-well pre-filled spin plate	Pre-filled spin columns	10 mL resin	96-well pre-filled spin plate
Resin bed volume(s)	0.1 mL	0.025–0.2 mL	0.1 mL	0.125 mL 0.5 mL 2 mL 4 mL	0.125 mL– 10 mL	0.55 mL
Cat. No.	88306	88305	88307	87777	87780	88304

Standard detergent removal resin results
HiPPR results

Table 1. Results using standard Thermo Scientific Pierce Detergent Removal Spin Column, 0.5 mL.

Detergent removal efficiency and protein recovery. BSA sample (25–200 µL) + detergent in 0.15 M NaCl, 0.05% sodium azide was mixed with equal volume of detergent removal resin (2x volume for CHAPS removal).

Detergent	Sample volume (µL)	Protein quantity (µg)	Detergent removal (%)	Protein recovery (%)
SDS (1%)	25	0.375	>99	98
	50	0.75	>99	97
	100	1.5	>99	100
	200	3.0	>99	100
Triton X-100 (1%)	25	0.375	>95	82
	50	0.75	>95	86
	100	1.5	>95	86
	200	3.0	>95	93
NP-40 (0.75%)	25	0.375	95	90
	50	0.75	96	94
	100	1.5	97	91
	200	3.0	97	97
CHAPS (1%)	25	0.375	95	64
	50	0.75	97	70
	100	1.5	98	78
	200	3.0	98	75

Results using standard Detergent Removal Spin Column

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Figure 1. Results using standard Pierce Detergent Removal Spin Column, 0.5 mL. Tryptic digests (0.1 mL, 100 μg) containing detergent were each processed through 0.5 mL of Pierce Detergent Removal Resin and subjected to LC-MS/MS analysis. Top row: Base peak LC-MS chromatograms. Bottom row: Integrated mass spectra. Similar results were produced for Brij-35 detergent, octyl glucoside, octyl thioglucoside, and SDS (data not shown).

Results using Detergent Removal Spin Columns

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Figure 2. Results using Pierce Detergent Removal Spin Columns, 0.5 mL. A tryptic digest of HeLa cell lysate (0.1 mL, 100 μg) containing 1% SDS was processed through 0.5 mL of Pierce Detergent Removal Resin and subjected to LC-MS/MS analysis. The processed sample allowed similar numbers of identified peptides as digests containing no SDS. Peptide identification is greatly reduced in sample containing SDS. Effective detergent removal enables greater peptide identification.

Table 2. Results using Thermo Scientific HiPPR Detergent Removal Resin.

Each column and plate well contained ~550 μL of detergent-removal resin slurry and 0.1 mL of sample.

Process format	Detergent	Detergent concentration (%)	Detergent removal (%)	BSA recovery (%)
0.5 mL Spin Column	Sodium deoxycholate	5	99	100
	Octyl glucoside	5	99	90
	Octyl thioglucoside	5	99	95
	Lauryl maltoside	1	98	99
	Triton X-114	2	95	100
	Brij-35	1	99	97
	Tween 20	0.25	99	87
96-well Spin Plate	SDS	5	99	89
	Triton X-100	4	99	100
	NP-40	1	95	100
	CHAPS	5	99	100

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Figure 3.Results using HiPPR Detergent Removal Resin. BSA (100 μg/mL) tryptic digests were prepared without detergent, in the presence of 0.5% Triton X-100 or spiked with 0.5% SDS following enzymatic digestion. Samples (0.1 mL) containing detergent were processed with the HiPPR Detergent Removal Resin and compared to unprocessed or detergent-free samples by LC-MS/MS. Results demonstrate that detergent removal is effective and produces results similar to those observed for samples containing no detergent.

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Figure 4. Results using HiPPR Detergent Removal Resin. BSA (25 and 100 μg/mL) was digested in the presence and absence of detergents and the samples were processed for LC-MS/MS analysis. Effective detergent removal resulted in greater peptide identification and high MASCOT™ scores.

Standard detergent removal resin results

Table 1. Results using standard Thermo Scientific Pierce Detergent Removal Spin Column, 0.5 mL.

Detergent	Sample volume (µL)	Protein quantity (µg)	Detergent removal (%)	Protein recovery (%)
SDS (1%)	25	0.375	>99	98
	50	0.75	>99	97
	100	1.5	>99	100
	200	3.0	>99	100
Triton X-100 (1%)	25	0.375	>95	82
	50	0.75	>95	86
	100	1.5	>95	86
	200	3.0	>95	93
NP-40 (0.75%)	25	0.375	95	90
	50	0.75	96	94
	100	1.5	97	91
	200	3.0	97	97
CHAPS (1%)	25	0.375	95	64
	50	0.75	97	70
	100	1.5	98	78
	200	3.0	98	75

Click image to enlarge

HiPPR results

Table 2. Results using Thermo Scientific HiPPR Detergent Removal Resin.

Each column and plate well contained ~550 μL of detergent-removal resin slurry and 0.1 mL of sample.

Process format	Detergent	Detergent concentration (%)	Detergent removal (%)	BSA recovery (%)
0.5 mL Spin Column	Sodium deoxycholate	5	99	100
	Octyl glucoside	5	99	90
	Octyl thioglucoside	5	99	95
	Lauryl maltoside	1	98	99
	Triton X-114	2	95	100
	Brij-35	1	99	97
	Tween 20	0.25	99	87
96-well Spin Plate	SDS	5	99	89
	Triton X-100	4	99	100
	NP-40	1	95	100
	CHAPS	5	99	100

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